Contemporarily, serial block-face scanning electron microscopy (SBF-SEM) has emerged as an immensely powerful nanoscopic imaging technique, capable of generating large-volume three-dimensional information on a variety of biological specimens in a semiautomated manner. Despite the plethora of insights and advantages provided by SBF-SEM, a major challenge inherent to the technique is that of electron charging, which ultimately reduces attainable resolution and detracts from overall image quality. In this chapter, we describe a pre-embedding approach that involves infiltration of tissue with a highly conductive silver filler suspension following primary fixation. Such an approach is demonstrated to improve overall sample conductivity, resulting in the minimization of charging under high-vacuum conditions and an improvement in lateral resolution and image contrast. The strength of this sample preparation approach for SBF-SEM is illustrated on liver tissue.
Shami, G. J., Cheng, D., & Braet, F. (2018). Silver filler pre-embedding to enhance resolution and contrast in multidimensional SEM: A nanoscale imaging study on liver tissue. In Methods in Molecular Biology (Vol. 1814, pp. 561–576). Humana Press Inc. https://doi.org/10.1007/978-1-4939-8591-3_33